Expression and promoter analysis of <i>Xenopus DMRT1</i> and functional characterization of the transactivation property of its protein

Yoshimoto, Shin; Okada, Ema; Oishi, Takeshi; Numagami, Ruriko; Umemoto, Hirohito; Tamura, Kei; Kanda, Hiromi; Shiba, Tadayoshi; Takamatsu, Nobuhiko; Ito, Michihiko

doi:10.1111/j.1440-169x.2006.00894.x

Cited by 34 publications

(30 citation statements)

References 29 publications

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“…The Nterminal region (amino acids 1-123), including the DNA-binding domain of DM-W, which is encoded by exons 2 and 3, has relatively high sequence identity (82%) with the corresponding region of DMRT1. By contrast, DM-W has no homologous region to exons 4-6 of DMRT1, which includes the sequence encoding the transactivation domain (Yoshimoto et al, 2006;Yoshimoto et al, 2008). The sequence of the DM-W exon 4, including an in-frame stop codon, contained no significant homology among genomic and cDNA sequences derived from vertebrate species, including X.…”

Section: Exogenous Dmrt1 Can Induce Primary Development Into Testes Imentioning

confidence: 99%

“…Transgenic tadpoles or frogs were produced using restriction-enzymemediated integration (REMI), as described previously (Yoshimoto et al, 2006). To detect the transgenes, PCR was performed using the genomic DNA from the liver of an individual frog or tadpole as the template, and specific primer pairs as follows: pcDNA3-FLAG-DMRT1, 5Ј-CAGCCAGGATTCTGGAGTTA-3Ј and 5Ј-CAACTAGAAGGC ACA -GTCG-3Ј; psh-DM-W1/2, 5Ј-CCCGTGTGGCAGATG ATT GTT ACCT-3Ј and 5Ј-TTTAGAAGGCACAGTCGAGG-3Ј.…”

Section: Production Of Transgenic X Laevismentioning

confidence: 99%

“…The Cterminal region of DMRT1 contains a transactivation domain (Yoshimoto et al, 2006), suggesting that DM-W functions as a competitor of DMRT1 for its target genes in the primordial ZW gonads. Interestingly, DM-W mRNA is more abundant than DMRT1 mRNA in the ZW gonads early in sex determination (Yoshimoto et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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Opposite roles ofDMRT1and its W-linked paralogue,DM-W, in sexual dimorphism ofXenopus laevis: implications of a ZZ/ZW-type sex-determining system

et al. 2010

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SUMMARYA Y-linked gene, DMY/dmrt1bY, in teleost fish medka and a Z-linked gene, DMRT1, in chicken are both required for male sex determination. We recently isolated a W-linked gene, DM-W, as a paralogue of DMRT1 in Xenopus laevis, which has a ZZ/ZW-type sex-determining system. The DNA-binding domain of DM-W shows high sequence identity with that of DMRT1, but DM-W has no significant sequence similarity with the transactivation domain of DMRT1. Here, we first show colocalization of DM-W and DMRT1 in the somatic cells surrounding primordial germ cells in ZW gonad during sex determination. We next examined characteristics of DM-W and DMRT1 as a transcription factor in vitro. DM-W and DMRT1 shared a DNA-binding sequence. Importantly, DM-W dose-dependently antagonized the transcriptional activity of DMRT1 on a DMRT1-driven luciferase reporter system in 293 cells. We also examined roles of DM-W or DMRT1 in gonadal formation. Some transgenic ZW tadpoles bearing a DM-W knockdown vector had gonads with a testicular structure, and two developed into frogs with testicular gonads. Ectopic DMRT1 induced primary testicular development in some ZW individuals. These observations indicated that DM-W and DMRT1 could have opposite functions in the sex determination. Our findings support a novel model for a ZZ/ZW-type system in which DM-W directs female sex as a sex-determining gene, by antagonizing DMRT1. Additionally, they suggest that DM-W diverged from DMRT1 as a dominant-negative type gene, i.e. as a 'neofunctionalization' gene for the ZZ/ZW-type system. Finally, we discuss a conserved role of DMRT1 in testis formation during vertebrate evolution.

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Section: Exogenous Dmrt1 Can Induce Primary Development Into Testes Imentioning

confidence: 99%

Section: Production Of Transgenic X Laevismentioning

confidence: 99%

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Opposite roles ofDMRT1and its W-linked paralogue,DM-W, in sexual dimorphism ofXenopus laevis: implications of a ZZ/ZW-type sex-determining system

et al. 2010

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“…MAB-3 acts as a transcriptional repressor, as does the male isoform of DSX, but the female isoform of DSX can activate transcription (9,14,16). A fusion of the C-terminal domain of Xenopus DMRT1 to the yeast GAL4 DNA-binding domain can activate reporter gene transcription in transfected cells, whereas in the fish Medaka Dmrt1a and its paralog Dmrt1bY/DMY can repress reporter transcription in transfected cells (17,18). Finding the in vivo targets of DMRT1 and determining how their expression is regulated are critical to a better understanding of how DMRT1 directs testicular development and function.…”

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confidence: 99%

Genome-wide analysis of DNA binding and transcriptional regulation by the mammalian Doublesex homolog DMRT1 in the juvenile testis

Murphy

Sarver²,

Rice

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

103

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The DM domain proteins Doublesex-and MAB-3-related transcription factors (DMRTs) are widely conserved in metazoan sex determination and sexual differentiation. One of these proteins, DMRT1, plays diverse and essential roles in development of the vertebrate testis. In mammals DMRT1 is expressed and required in both germ cells and their supporting Sertoli cells. Despite its critical role in testicular development, little is known about how DMRT1 functions as a transcription factor or what genes it binds and regulates. We combined ChIP methods with conditional gene targeting and mRNA expression analysis and identified almost 1,400 promoter-proximal regions bound by DMRT1 in the juvenile mouse testis and determined how expression of the associated mRNAs is affected when Dmrt1 is selectively mutated in germ cells or Sertoli cells. These analyses revealed that DMRT1 is a bifunctional transcriptional regulator, activating some genes and repressing others. ChIP analysis using conditional mutant testes showed that DNA binding and transcriptional regulation of individual target genes can differ between germ cells and Sertoli cells. Genes bound by DMRT1 in vivo were enriched for a motif closely resembling the sequence DMRT1 prefers in vitro. Differential response of genes to loss of DMRT1 corresponded to differences in the enriched motif, suggesting that other transacting factors may modulate DMRT1 activity. DMRT1 bound its own promoter and those of six other Dmrt genes, indicating auto-and cross-regulation of these genes. Many of the DMRT1 target genes identified here are known to be important for a variety of functions in testicular development; the others are candidates for further investigation.roteins related to Doublesex (DSX) of Drosophila control sex determination and sexual differentiation in a broad array of metazoan animals and comprise the only family of deeply conserved sexual regulators so far identified (1). These proteins share the DM domain, a highly intertwined zinc finger DNA-binding motif first identified in DSX and the related Caenorhabditis elegans sexual regulator MAB-3 (male abnormal 3) (2-4). The vertebrate DM domain gene Dmrt1 and its close orthologs act as primary sex-determining genes in vertebrate clades including fish, amphibians, and probably birds, each with an independently evolved chromosomal sex determination mechanism (5-7). Thus Dmrt1 homologs frequently are recruited or retained to determine sex as new sex determination mechanisms arise. The targets of DM domain gene regulation also have important evolutionary roles: In insects the evolution of cis-regulatory elements bound by DSX appears to have driven divergence of sex-specific characters and thus may help drive speciation (8,9).Functional studies in the mouse have demonstrated that Doublesex and MAB-3 related transcription factor 1 (DMRT1) is essential for testicular differentiation (10). In mice DMRT1 is expressed in premeiotic germ cells and in Sertoli cells (the somatic "nurse" cells for the male germ line). Cell type-specific ge...

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“…Dmrt1 was the only gene whose expression was restricted to the gonadal tissue in both male and female, and the Bidder's organ. This gonad specific expression of Dmrt1 is also observed in X. laevis, R. rugosa, as well as in other taxa Yoshimoto et al, 2006;Lei et al, 2007). The conserved gonad-specific expression pattern in many vertebrate lineages reflects its ancient association with sex-determination and sexual differentiation within both invertebrates and vertebrates (Raymond et al, 1998.…”

Section: Discussionmentioning

confidence: 81%

Cloning and expression of candidate sexual development genes in the cane toad (Bufo marinus)

Abramyan

Feng

Koopman

2009

Developmental Dynamics

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The development of the reproductive system in bufonids (true toads) is unique in several respects: sexual differentiation occurs later than in other anurans, and toads develop a Bidder's organ, a rudimentary ovary that can be manipulated in males to produce mature oocytes. To illuminate the genesis of this unusual reproductive system, we isolated from the cane toad (Bufo marinus) the orthologues of several known vertebrate sex-determining genes, determined their primary structure, and studied their expression by reverse transcriptase-polymerase chain reaction and in situ hybridization of tissue sections. We report here that cane toad Sox9, Dmrt1, and p450aromatase (Cyp19a1) are highly homologous to their counterparts in other vertebrates. They show profiles of expression that generally follow patterns observed in other taxa, but with some novel features. Our data suggest that these genes likely play key roles in sex determination and early gonad development in bufonids.

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Expression and promoter analysis of Xenopus DMRT1 and functional characterization of the transactivation property of its protein

Cited by 34 publications

References 29 publications

Opposite roles ofDMRT1and its W-linked paralogue,DM-W, in sexual dimorphism ofXenopus laevis: implications of a ZZ/ZW-type sex-determining system

Opposite roles ofDMRT1and its W-linked paralogue,DM-W, in sexual dimorphism ofXenopus laevis: implications of a ZZ/ZW-type sex-determining system

Genome-wide analysis of DNA binding and transcriptional regulation by the mammalian Doublesex homolog DMRT1 in the juvenile testis

Cloning and expression of candidate sexual development genes in the cane toad (Bufo marinus)

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